BACKGROUND: Vascular endothelial growth factor receptor 2 (VEGFR2) is primarily involved in vascular endothelial growth factor (VEGF)-mediated signal transduction and plays a critical role in the pathological angiogenesis that occurs in a number of diseases, including leukemia. Besides, VEGF secreted by leukemia cells also induces its own expression which leads to an enhanced production of VEGFR2 which contributes to the survival and proliferation of leukemia cells.OBJECTrVE: To evaluate the inhibitive effect of Lenti6/shVEGFR2 on the VEGFR2 expression and leukemia growth in mouse.DESIGN: A randomized, parallelized, controlled and open trial.SETTING: Department of Pediatrics, the Second Affiliated Hospital of Sun Yat-sen University; Biotechnology Research Center, Sun Yat-sen University.MATERIALS: The experiment had been done in the laboratories for Medical Research Center of the Second Affiliated Hospital, Sun Yat-sen University and Biotechnology Research Center, Sun Yat-sen University from May 2004 to January Lentiviral RNAi Expression System was purchased from Invitrogen, Co.,Ltd.; human VEGFR2 Mcb (PE) was purchased from R&D; CD31 immunohistochemistry kit was purchased from Boster, Co.,Ltd.; CD33-PE fluorescence labeled antibody was purchased from BD, Co.,Ltd.transiently and expression clone (Lenti6/shVEGFR2) was constructed, then cotransfected with ViraPowerTM Packaging Mix pU6/shVEGFR2 entry clone and transducting with Lenti6/shVEGFR2 expression clone, the effect on the development of intravenous xenograft leukemia mouse model, the distribution of microvessels in mouse bone marrow was observed after leukemia model mouse injected with recombinant lentivirus (group B); leukemia model mouse injected with recombinant lentivirus and endothelial cell (group C); leukemia model mouse injected with endothelial cell (group D). Through detecting changes of CD33 positive cells and microvessel density (MVD) in bone marrow, observing peripheral blood cell (PBC)smear and slice of liver, spleen, the effect of Lenti6/shVEGFR2 recombinant lentivirus on mouse leukemia was evaluated.mediated with lentivirus on VEGFNEGFR2 paracrine and autocrine loops in leukemia mouse.effective in inhibiting HL60 cell. pU6/shVEGFR2 entry clone constructed according to it had cell inhibitory rate as high as after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone: 48 hours after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone, the cell growth inhibitive rates were similar. However, the cell growth inhibitive rate of entry clone descended rapidly after 48 hours (P＜0.01); which of expression clone changed slowly, reaching the peak at 96 hours, dropped slightly, having no significance mouse: The amount of HL60 cells in bone marrow of groups A, B and C detected with flow cytometry were (25.8%±4.9)%, (14.3%±5.1)%, (8.4±2.6)%, respectively (P＜0.05); MVD in group C was obviously less than that in group D (P＜0.05); The amount of HL60 cells in leukemia model mouse injected with recombinant lentivirus and endothelial cell was the lowest as compared with the other groups.

To establish a scientific and feasible evaluation index system and to assess medical research programs subsidized by the Shanghai Municipal Health Bureau in 2002-2006, so as to provide reference to aid the management department in decision-making and better guidance of research projects. Delphi expert consultation was employed to construct an evaluation index system, and used the system to comprehensively assess the funded projects. In the end, we found that the funding of the Shanghai Health Bureau had certain promoting effect in optimization of research team, startup of scientific research, training of research personnel, and the development of disciplines.

Objective To investigate the optimal dose of apocynin to protect severe acute pancreatitis (SAP) and SAP caused intestinal injury in rats. Methods A total of 53 SPF male Wistar rats were randomly allocated into five groups:sham operation group (SO group, n=10), SAP group (n=12), low-dose apocynin group (25 mg/kg,n=11), medium-dose apocynin group (50 mg/kg, n=10) and high-dose apocynin group (100 mg/kg,n=10). SAP model was prepared by retrograde infusing 5%sodium taurocholate (1 mL/kg) into biliopancreatic duct of rat. At thirty minutes before modeling, apocynin was injected into rat to intervention. The survival condition was recorded at 12 h after modeling, and blood samples were obtained for detecting serum amylase (AMY), alanine aminotransferase (ALT) and creatinine (Cr). Pancreatic and ileal tissue samples were obtained for HE staining and pathological examination. Results Two rats died in SAP group and one died in low-dose apocynin group. The quantity of ascites, the levels of AMY, ALT, Cr and pancreatic and intestinal pathologic scores were significantly increased in SAP group than those in SO group (P<0.05). Except the levels of Cr and intestinal pathologic score, there was no significant difference between low-dose apocynin group and SAP group. The quantity of ascites ascites, levels of AMY, Cr and pancreatic and intestinal pathologic scores were significantly lower in medium-dose and high-dose apocynin groups than those in SAP group (P<0.05). The levels of ALT and Cr were significantly higher in high-dose apocynin group than those of medium-dose apocynin group (P<0.05). Conclusion Apocynin improves SAP symptoms and reduces SAP caused intestinal injury in rats, which may be related to the inhibition of NOX activity, and 50 mg/kg of apocynin is the optimal dose.

Objective To evaluate the curative effect of infection caused by peritoneal dialysis(PD) catheter. Methods Eighty patients with infections caused by peritoneal dialysis catheter were enrolled in this study and randomized into silver-containing hydrofiber dressing group and control group with 40 patients in each group. The control group was treated with iodophor and/or gentamicin and the treatment group with silver-containing hydrofiber dressings. The two groups were compared in terms of curative effect at the wounds and time for cure. Result The cure rate in the treatment group was significantly higher than that in the control group (P<0.05), and the cure time in the observation group was significantly shorter as compared to the control group. Conclusion Silver-containing hydrofiber dressing is effective in the treatment of PD catheter-induced infections , and it can shorten treatment duration and reduce the frequency of dressing change.

Objective To investigate the role of titanium plate fixation for sternum closure after off-pump coronary artery bypass grafting (OPCABG) in elderly patients.Methods A total of 120 elderly patients who accepted OPCABG were randomly divided into two groups.The control group (n =60) received wire to fix the sternum,and the experimental group (n=60) received titanium plate combined with wire to fix the sternum.The chest closure time,the rate of postoperative incision pain,the rate of sternal dehiscence and sternal re-fixation between two groups were observed and recorded.Results The rates of postoperative incision pain,sternal dehiscence,sternal re-fixation was lower in experimental group than in control group [11.7％(7/60) vs.38.3％(23/60),3.3％(7/60) vs.13.3％(8/60),1.7％(1/60) vs.11.7％(7/60),x2 =11.378,3.927,4.821 respectively all P＜0.05].The chest closure time was longer in experimental group than in control group [(39±5)min vs.(30±2) min,x2 =13.386,P＜0.05].Conclusions Titanium fixation plate combined with wire for sternum closure can increase the stability of sternum fixation,reduce the postoperative pain and sternal dehiscence,and then decrease the risk of sternal re-fixation,but delay sternal closure of OPCABG in the elderly.

Objective To explore the effect of pleural cavity integrity on respiratory system after off-pump coronary artery bypass grafting (OPCABG),through comparing the respiratory complication after OPCABG.Methods One hundred and two patients were accepted OPCABG,among whom 49 patients' pleural cavities were opened (open group) and 53 patients' pleural cavities were closed (close group).The ventilation time,intensive care unit time,pleural effusion,the rate of atelectasis and respiratory failure after operation were compared between two groups.Results The ventilation time and intensive care unit time in open group were (40.3 ± 4.8) h and (78.3 ± 10.8) h,in open group were (28.6 ± 6.8) h and (54.8 ± 6.1) h.The ventilation time and intensive care time in open group were significantly longer than those in close group(P ＜ 0.01 or ＜ 0.05).The pleural effusion in open group was (800.0 ± 60.5) ml,in close group was (350.0 ± 28.6) ml.The pleural effusion in open group was significantly higher than that in close group (P ＜ 0.01).The rate of postoperative atelectasis and respiratory failure in open group were 36.7％(18/49) and 38.8％(19/49),in close group were 15.1％(8/53) and 18.9％(10/53).The rate of postoperative atelectasis and respiratory failure in open group were significantly higher than those in close group (P ＜ 0.01).Conclusions OPCABG is the operation in mediastinum.To avoid pleural cavity opened in OPCABG can reduce the incidence of postoperative respiratory complication.

Objective To evaluate the effectiveness of low-dose erythromycin for the treatment of feeding intolerance in preterm infants in China.Methods In this study,random clinical trials on the treatment of feeding intolerance in preterm infants with intravenous low-dose erythromycin published were searched at Chinese Journal Full-text Database,Chongqing Weipu Database and Wanfang database by using the methods of Cochrane systematic review.At the same time the information from related journals,professional data and network were hand-searched.The publishing deadline for the literatures reviewed in this study was August 2012.Statistical analysis of clinical data was performed by using RevMan 4.2 software provided by the Cochrane Collaboration.Results A total of 9 studies were included.The results showed that compared with the group of comprehensive therapy,the group of low-dose erythromycin was superior in the following aspects with significant differences(P ＜ 0.05):the average length of hospital stay,time of parenteral nutrition,time to full feeding,the incidence rate of feeding intolerance (Z =3.44,P =0.000 6 ; Z =6.78,P ＜0.000 01 ; Z =3.96,P ＜ 0.000 1 ; Z =2.51,P =0.01).Conclusion Low-dose erythromycin therapy for feeding intolerance in preterm infants is superior to the comprehensive therapy.It provides a prospective therapeutic method for feeding intolerance in preterm infants.However,large scale,multicenter and well-designed clinical trials should be adopted to confirm the conclusions.

Objective To evaluate the consistency of the test results detected by multiple automated hematology analyzers .Meth-ods The EP9-A2 guideline of NCCLS ,Passing-Bablok regression analysis and t test were adopted to compare the detection results of hemoglobin(HGB) ,hematocrit (HCT) ,red blood cell count(RBC) ,white blood cell count (WBC) and platelet(PLT) in 115 samples from patients by 5 automated haematology analyzers of the Sysmex XE-2100 ,the Sysmex XT-1800i ,the Sysmex XS-1000i , the Mindray BC-5300 and ABX pentra80 .The Sysmex XE-2100 was used as the reference instrument .Results In the detection of all the compared items including HGB ,HCT ,RBC ,WBC and PLT ,these automated hematology analyzers exhibited very good cor-relation(r>0 .97) ,whereas the individual parameters in 3 automated hematology analyzers appeared the proportional systematic differences and /or constant systematic differences ,including HCT in the sysmex XT-1800i ,HCT and RBC in the sysmex XS-1000i and RBC in the ABX pentra 80 .Conclusion The Passing-Bablok regression analysis combined with t test may identify the system-atic differences of the comparative results of 2 automated haematology analyzers ,the consistency of the comparative results of the automated hematology analyzers can not be evaluated by the simple correlation coefficient .

BACKGROUND: Previous studies have confirmed that embryonic stem cells call be induced to differentiate into insulin-producing cells, but the induction process takes a long time. Most of the processes take about one month.OBJECTIVE: Activin A, all-trans retinoic acid (ATRA), basic fibroblast growth factor (bFGF) and nicotinamide were applied in vitro in combination to observe whether mouse embryonic stem cells could be induct to differentiate into insulin-producing cell in a relatively short time.DESIGN: Cell observation experiment.SETTING: Shanghai Institute of Endocrine and Metabolic Diseases, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.MATERIALS: This study was performed at Shanghai Institute of Endocrine and Metabolic Diseases from October 2004 to February 2006. Two mice of clean grade and of 12.5-14.5 days of gestational age were provided by Shanghai SLAC Laboratory Animal Co., Ltd (Permission No. 2004A034). The protocol was performed in accordance with ethical guidelines for the use and care of animals. Mouse embryonic stem cell lines were supplied by Dr Changxian Zhang (CNRS UMR5641, France). Activin A was the product of the R & D Corporation. ATRA and nicotinamide were supplied by the Sigma Corporation, USA. BFGF was supplied by Gibco Corporaion. METHODS: Head and viscera were removed from embryos of the pregnant mouse. The remaining tissues were cut into pieces and digested with trypsin. Cell suspension was centrifuged and inoculated at 3×108L-1. The cells could be used as mouse feeder layer after 2-3 times of passage. The mouse embryonic stem cells (ESCs) were inoculated onto the feeder layer in knockout Dulbecco's modified Eagle medium (DMEM) supplemented with leukemia inhibitory factors (LIF). ESCs were passaged at 1:3-1:6 after 2-3 days of culture. Culture medium with serum was added into the culture dishes to terminate the digestion. Cell fluid was centrifuged and supernatant was discarded. The sediments were prepared into suspension and inoculated at 2.5×104 with LIF-free culture medium. After 24-48 hours, embryonic bodies (EBs) were collected and replated in 1% Matrigel-coated dishes. When began to adhere to the dishes, EBs were cultured in 10% FBS/DMEM supplemented with 100μg/L activin A for 24 hours. Then EBs were switched to 10% FBS/DMEM for 6-8 hours as an interval. After this interval. EBs differentiated were cultured in 10% FBS/DMEM with 10<-6mol/L RA for another 24 hours followed by culture in 10% FBS/DMEM supplemented with 10μg/L bFGFs for 3-5 days. Finally, EBs differentiated were cultured in DMEM/F12 supplemented with N2 supplement, B27 supplement, 1μg/L laminin, 10μg/L bFGFs, and 10mmol/L nicotinamide for 3-5 days. Dithizone (DTZ) staining, inununofluorescent staining and reverse transcription-polymerase chain reaction (RT-PCR) were applied to detect insulin expression in the differentiated cells.MAIN OUTCOME MEASURES: Induction of ESCs, DTZ staining and immunofluorescent staining as wel as RT-PCR detection.RESULTS: Mouse ESCs growing on a feeder layer formed many colonies with clear boundary and dense structure. However, there was no obvious outer limit between these ESCs. EBs began to adhere to the dishes, which were coated with matrigel, on the 2nd day. After activin A and ATRA interval induction, EBs spread, and most of the living cells were epithelial cell-like when cultured in 10% FBS/DMEM supplemented with 10μg/L bFGFs. After culturing in DMEM/F12 supplemented with N2, B27, nicotinamide, bFGFs and laminin, the cells formed small clusters. The insulin-producing cells were stained dark red with DTZ, and the cells stained with primary antibody to insulin were insulin-positive. After 2 weeks of induction of activin A, ATRA, bFGFs and nicotinamide, the insulin-producing cells expressed insulin 2, Pdxl, Nkx6.1, Nkx2.2, PP, IAPP, Glut2, Somastatin, Hnf3β and Neuro D mRNA but did not express insulin 1 mRNA.CONCLUSION: Mouse ESCs call be induced to differentiate into insulin-producing cells by activin A, ATRA, bFGFs and nicotinamide in vitro. Induction time call be shortened to 2 weeks.

The methods of 1H Nuclear Magnetic Resonance (NMR) and mass spectroscopy were used in detecting the metabolites of brodimoprim (BDP) in rat plasma. The endogenic compounds in the plasma were removed with solid phase extraction (SPE) column firstly, then the mixture of metabolites was identified with 1H NMR and mass spectroscopy (MS). Two metabolites of BDP in the plasma at 20h were detected, they were demethyl-brodimorpim glucuronide and brodimoprim sulfurate. The study proved that the method of SPE coupled with NMR and MS can be applied to the analysis of metbolites in plasma quickly and conveniently.